Postersot2013_niehof

Cytochrome P450 monooxygenases expression in human epithelial lung cell linesAbbreviationConcentrationReceptorCalu-3 cells, CYP gene expression
Fraunhofer Institute for Toxicology and Experimental Medicine, Hannover, Germany
Table 1: Various CYP inducers used for the treatment of Calu-3 and A549 cells.
INTRODUCTION2C(8-19)
The pulmonary epithelium is the first barrier for airborne
Basal expression of CYP1B1, CYP1A1, CYP2J2, CYP2E1,
monooxygenases (CYP) participate in metabolic inacti-
CYP3A7 was detected in both cell lines (Fig. 1, Fig, 2),
vation of xenobiotics. Beyond that some compounds
which is consistent with CYP expression in the human
Ct-values (threshold cycles), real time RT-qPCR2C(8-19)
require enzymatic activation to exert their toxic effects or
lung. Furthermore, potential CYP inducers were analyzed
their desirable functions. The bronchiale epithelial cell line
(Table 1). Omeprazole acts on aryl hydrocarbon receptor
Figure 1: CYP expression in untreated Calu-3 cells 48 h after seeding
Calu-3 and the type II-like pulmonary epithelial cell line
(AhR) activation and induced CYP1A1 and CYP1B1 in
(n = 3). Gene expression was analysed by real time RT-qPCR. Ct
A549 serve as cell culture models for the human
both cell lines (Table 2, 3). Rifampicin acts on pregnane x
(threshold cycle) reflects the cycle number at which the fluorescence
generated within a reaction crosses the threshold.
respiratory epithelium. So far, there is little information
receptor (PXR) and phenobarbitale acts predominantly on
regarding their metabolic properties especially for Calu-3
constitutive androstane receptor (CAR), however, both
Table 2: Fold change in CYP gene expression in Calu-3 cells 24 h after
cells. In this study we want to further characterize A549
receptors are not expressed in the lung. Accordingly, both
treatment with different inducers. Fold changes > 2.5 with p<0.05 (n = 3) are
A549 cells, CYP gene expression
and Calu-3 cells regarding their basal and inducible CYP
agents did not induce any CYP in these cells (Table 2,
indicated in bold (green boxes). Analysis was performed with the opensource software REST 2009 (TU Munich/Qiagen) using multiple different
reference genes, respectively. nd = not detected.
members of the CYP3A family, mainly CYP3A7 in Calu-3
cells (Table 2), and CYP3A5 and CYP3A7 in A549 cells
MATERIAL AND METHODS
(Table 3). CITCO is known to act as a CAR agonist and is
normally used to induce CYP2B6/7. However, it is a potent
inducer of CYP1B1 in Calu-3 cells (Table 2), and of
Cells were cultured according to ATCC conditions. After
CYP1A1 and CYP1B1 in A549 cells (Table 3).2C(8-19)
24 h cells were treated with the respective inducers or
2C(8-19)
left untreated. Again after 24 h cells were harvested and
RNA was isolated. Subsequently, CYP expression was
CONCLUSION
quantitative polymerase chain reaction (RT-qPCR) with
Ct-values (threshold cycles), real time RT-qPCR
the Light Cycler SystemTM (Roche) or ABI Prism® 7500
Calu-3 cells and A549 cells express a broad range of
Table 3: Fold change in CYP gene expression in A549
CYPs with preserved inducibility and are valuable models
Figure 2: CYP expression in untreated A549 cells 48 h after seeding
treatment with different inducers. Fold changes > 2.5 with p<0.05 (n = 3) are
(n = 3). Gene expression was analysed by real time RT-qPCR. Ct
indicated in bold (green boxes). Analysis was performed with the open
of the airway epithelial barrier for metabolic in vitro
(threshold cycle) reflects the cycle number at which the fluorescence
source software REST 2009 (TU Munich/Qiagen) using multiple different
generated within a reaction crosses the threshold.
reference genes, respectively. nd = not detected.CONTACT [email protected]For further information please also visit us at exhibition booth # 1449